免疫荧光技术在鱼类生殖细胞发育研究中的应用  

Application of Immunofluorescence Technique in Studying Development of Fish Germ Cells

作　　者：杨聪慧 杨福忠 朱辣 罗心玥 赵如榕[1] 张纯[1] YANG Conghui;YANG Fuzhong;ZHU La;LUO Xinyue;ZHAO Rurong;ZHANG Chun(State Key Laboratory of Developmental Biology of Freshwater Fish,College of Life Science,Hunan Normal University,Changsha 410081,China;Hunan Xiangyun Biotechnology Limited Company,Changde 415101,China)

机构地区：[1]省部共建淡水鱼类发育生物学国家重点实验室,湖南师范大学生命科学学院,长沙410081 [2]湖南湘云生物科技有限公司,常德415101

出　　处：《激光生物学报》2022年第5期411-416,共6页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(31873038);湖南省自然科学基金青年项目(2020JJ5357);湖南省教育厅重点项目(20A289)。

摘　　要：在生殖细胞发育的相关研究中,功能分子标记在人类及其他模式生物中已经得到了广泛应用,但在鱼类中应用较少。本文采用RNA PolⅡCTD(S5)作为功能分子标记对鲤雄性生殖细胞的细胞核进行了免疫荧光研究,并对相关技术参数进行了深入的探索。研究结果表明,该方法可以在鲤雄性生殖细胞的多个发育阶段检测到与转录活性位点相关的染色体免疫荧光信号及细胞相特征,并发现荧光信号的质量会受到染色体制片及洗片等关键步骤的影响。本研究提供了一个重要的功能分子标记,并将之作为免疫探针在经济鱼类生殖细胞发育研究中进行尝试性应用,这为鱼类遗传育种中发育功能的检测提供了一个新的策略。Functional molecular markers have been widely used in the study of germ cell development of human and other model organisms,whereas less applications were in fishes.In this study,RNA PolⅡCTD(S5)was used as a functional molecular marker for immunofluorescence analysis in germ cell nuclei of common carp,and the related technical parameters were further explored.The results showed that this method could detect the characteristics of mitosis phase and chromosomal immunofluorescence signals,which were related to transcriptional active sites during the development stage of fish germ cells.In addition,the quality of immunofluorescence signals were affected by the critical experimental procedures including chromosome preparation and washing specimens.This study not only provides important molecular markers served as immunoprobe for the study of germ cell development in fishes,but also provides a new detection strategy for fish genetics and breeding.

关 键 词：鲤 生殖细胞 免疫FISH RNA聚合酶ⅡCTD(S5)蛋白 减数分裂 

分 类 号：S917.4[农业科学—水产科学]